Toll Like Receptor Response and traditional risk factors for atherosclerosis: Obesity is related to a higher inflammatory responsive state of circulating cells after stimulation of toll like receptor 2 and 4. V.P.W. Scholtes Experimental Laboratory Cardiology University Medical Center Utrecht The Netherlands ESC Congress, Stockholm 21 Adipose tissue and inflammation
Atherosclerosis and inflammation Adapted from Obesity, inflammation, and atherosclerosis Viviane Z. Rocha and Peter Libby Nat. Rev. Cardiol. 6, 399 49 (29)
Toll Like Receptor and inflammation Ligand Toll Like Receptor TNF-α IL-6 NF-κB DNA transcription
Toll like Receptor Response 1 μl of blood + 1 μl of LPS 1 μl of blood + 1 μl of P3C TLR4 monocyte monocyte TLR2 cytokines ELISA: TNF-alpha
TLR response: clinical perspective (1) Enhanced response of blood monocytes to in vitro LPS challenge in patients with recurrent unstable angina, Liuzzo Circulation 21;13;2236-2241
TLR response: clinical perspective (2) Monocyte toll-like receptor 2 and 4 responses and expression following PCI. Asocation with lesion stenosis and fractional flow reserve D Versteeg Heart 28 94: 77-776
Research question Are traditional risk factors and obesity associated with an altered Toll Like Receptor response?
Obesity and inflammation Inflammatory disorder Adapted from Obesity, inflammation, and atherosclerosis Viviane Z. Rocha and Peter Libby Nat. Rev. Cardiol. 6, 399 49 (29)
Methods Cohort 1 (PCI cohort) 1 Patients scheduled for Percutaneous Coronary Intervention Cohort 2 (CEA cohort) 113 Patients scheduled for Carotid Artery Endarterectomy Exclusion of: patients with an acute Myocardial infarction patients with a chronic inflammatory disease (corticosteroid use)
Clinical Characteristics PCI cohort CEA cohort parameter 78 patients 15 patients sex / 56 / 22 73 / 35 age median [IQR] 62,5 [53 7] 7,7 [63 78] BMI median [IQR] 26.9 [24.6 34.9] 26.4 [24.2 29.5] BMI > 25 n (%) 41(73%) 14(64%) 45(66%) 23(68%) smoking n (%) 5 (6%) 27 (25%) hypertension n (%) 52 (67%) 88 (84%) diabetes n (%) 16 (21%) 21 (2%) hypercholesteremia n (%) 72 (92%) 84 (8%) family history of CVD n (%) 41 (53%) 28 (27%)
TNF-alpha ng/ml TNF-alpha ng/ml Risk factors and TLR response CEA cohort PCI cohort 1 8 TLR 2 1 8 6 6 4 4 2 2 yes (52) no (26) yes (88) no (17) yes (16) no (62) yes (21) no (84) Hypertension Diabetes
TNF-alpha ng/ml TNF-alpha ng/ml Risk factors and TLR response PCI cohort CEA cohort 1 8 TLR 2 1 8 6 6 4 4 2 2 yes (72) no (6) yes (84) no (21) Hypercholesterolemia yes (41) no (37) yes (28) no (59) Family History of CVD
TNF-alpha pg/ml TNF-alpha pg/ml Smoking and TLR response PCI cohort CEA cohort 1 TLR 2 1 p =.13 TLR 4 5 5 no (73) yes (5) no (79) yes (26) no (73) yes (5) no (79) yes (26)
TNF-alpha pg/ml TNF-alpha pg/ml BMI and TLR response PCI cohort CEA cohort p =.4 TLR 2 1 p =.41 TLR 4 1 p =.254 p =.17 5 5 < 25 (37) > 25 (68) < 25 (23) > 25 (55) < 25 (37) > 25 (68) < 25 (23) > 25 (55)
cd11b expression (mean fluorescence) cd11b expression (mean fluorescence) BMI and inflammation: Cd11b expression on monocytes 8 Study 1 p =.6 3 p =.72 Study 2 6 2 4 2 1 cd11b < 25 (13) 25-3 (21) > 3 (9) BMI group cd11b < 25 (24) 25-3 (29) > 3 (11) BMI group
hscrp serum level mg/l BMI and inflammation: hscrp levels (study 1) 15 Study 1 p =.4 1 5 < 25 (23) 25-3 (31) > 3 (24) hscrp BMI group
Conclusion Obesity was the only risk factor for CVD that was positively associated with an altered TLR response We observed a trend towards lower levels of TNF-alpha for patients who were active smokers The enhanced systemic inflammatory state in obese patients was also reflected in: - elevated levels of hscrp - and increased expression of the activation marker cd11b on monocytes.
Acknowledgements Experimental Laboratory Cardiology Prof. G. Pasterkamp Prof DPV de Kleijn dr I Hoefer dr JK van Keulen dr DIK Versteeg EHAM Elsenberg Department of Cardiology Prof DA Doevedans dr PR Stella Department of Vascular Surgery Prof FL Moll dr GJ de Borst dr J.A. van Herwaarden dr R.J. Toorop Department of Vascular Surgery dr JPPM de Vries dr H.D.W.M. Van de Pavoordt dr J. Wille drs R.H. van de Mortel dr B.A.N. Verhoeven Department of Microbiology dr G.T. Rijkers mail: vscholt2@umcutrecht.nl
Extra figures
Smoking and Attenuation of TLR response Tobacco Smoking Inhibits Expression of Proinflammatory Cytokines and Activation of IL-1R-Associated Kinase, p38, and NF- {kappa}b in Alveolar Macrophages Stimulated with TLR2 and TLR4 Agonists Haiyan Chen, Mark J. Cowan, Jeffrey D. Hasday, Stefanie N. Vogel and Andrei E. Medvedev J. Immunol. 27;179;697-616
cd11b expression (mean fluorescence) cd11b expression (mean fluorescence) Very obese patients and lower TLR responsiveness 8 Study 1 p =.6 3 p =.72 Study 2 6 2 4 2 1 cd11b < 25 (13) 25-3 (21) > 3 (9) BMI group cd11b < 25 (24) 25-3 (29) > 3 (11) BMI group
TNF-alpha pg/ml TNF-alpha pg/ml TNF-alpha pg/ml TNF-alpha pg/ml Very obese patients have a lower TLR response 15 Study 1 6A p =.6 1 Study 1 6B p =.71 1 5 5 6 < 25 (23) 25-3 (31) > 3 (24) P3C5 (TLR2) Study 2 6C BMI group p =.6 6 < 25 (23) 25-3 (31) > 3 (24) LPS1 (TLR4) Study 2 6D BMI group p =.11 4 4 2 2 < 25 (37) 25-3 (44) > 3 (24) P3C5 (TLR2) BMI group < 25 (37) 25-3 (44) > 3 (24) LPS1 BMI group
TNF-alpha pg/ml TNF-alpha pg/ml BMI and TLR response 2 study 1 1 study 2 15 1 5 5 2 25 3 35 4 BMI 2 25 3 35 4 BMI
PCI cohort CEA cohort TLR2 stimulation present not present present not present Hypertension (26) 11 [3 35] (52) 18 [8 5] (88) 12 [6 22] (17) 14 [7 25] Diabetes (16) 1 [ 26] (62) 14 [6 44] (21) 12 [7 2] (84) 12 [6 25] Hypercholesterolemia (72) 12 [4 42] (6) 23 [8 4] (84) 11 [6 21] (21) 15 [9 32] Family history of heart disease < 6 years (41) 15 [6 46] (37) 11 [3 36] (28) 11 [7 32] (59) 12 [6 19] BMI ( BMI < 25 / BMI > 25) (55) 17 [8 49] (23) 7 [2 19] (37) 9 [6 16] (68) 14 [7 28] Smoking (yes / no) (5) 8 [2 22] (73) 13 [5 43] (26) 11 [6 2] (79) 12 [7 25] TLR4 stimulation present not present present not present Hypertension (26) 355 [178 565] (52) 246 [18 582] (88) 173 [99-296] (17) 245 [152-384] Diabetes (16) 246 [155 347] (62) 363 [182 587] (21) 196 [112 271] (84) 185 [13 315] Hypercholesterolemia (72) 36 [178 561] (6) 35 [27 694] (84) 185 [13 3] (21) 22 [146 329] Family history of heart disease < 6 years (41) 273 [177 555] (37) 359 [22 578] (28) 216 [95 336] (59) 165 [18 297] BMI ( BMI < 25 / BMI > 25) (55) 364 [196 599] (23) 219 [175 362] (37) 154 [15 261] (68) 29 [112 336] Smoking (yes / no) (5) 15 [7 231] (73) 353 [191 573] (26) 26 [125-312] (79) 181 [18-3]